Methods and devices for analysis of sealed containers

ABSTRACT

This invention provides methods, NMR probes, and NMR systems for the analysis of the contents of sealed food and beverage containers and the like.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.11/016,305 filed Dec. 16, 2004, which is a continuation of U.S. patentapplication U.S. Ser. No. 10/622,008 filed Jul. 16, 2003, now U.S. Pat.No. 6,911,822, which claims priority to and benefit of U.S. applicationSer. Nos. 60/396,644, filed Jul. 17, 2002 and 60/465,644, filed Apr. 25,2003, the full disclosures of which are incorporated herein byreference.

FIELD OF THE INVENTION

This invention relates to methods and devices for analyzing sealed foodand beverage containers, and particularly sealed wine bottles, by NMRspectroscopy.

BACKGROUND OF THE INVENTION

Wine is the product of the growth and metabolism of yeasts and bacteriain grape must. It is well known that many of these and other bacteriasurvive all of the steps of wine making from the mature grape throughvinification to bottle corking (Ribéreau-Gayon (1985) “New developmentsin wine microbiology” Am. J. Enol. Vitic. 36:1-10). One class oforganisms of interest is Acetobacter, a bacteria responsible foroxidizing ethyl alcohol into vinegar or acetic acid (Drysdale and Fleet(1988) “Acetic acid bacteria in winemaking: a review” Am. J. Enol.Vitic. 39:143-154; Millet and Lonvaud-Funel(2000) “The viable butnon-culturable state of wine micro-organisms during storage” Lt. Appl.Microbiol. 30:136-141). Although present in most wines, Acetobacter doesnot typically generate enough acetic acid to spoil wine during bottlestorage due to a lack of oxygen. As long as the wine is stored in ananaerobic environment, conditions ensured by quality corking, acceptablylow quantities of acetic acid (e.g., below sensory levels) are producedand the quality of the wine is preserved. Unfortunately, the sealingperformance of wine corks can degrade with age, and the long termbehavior of low quality natural corks and synthetic stoppers is not welldocumented. One consequence of a leaky cork is the admission of oxygento wine, a triggering of Acetobacter function, and the production ofacetic acid . Furthermore, the admission of oxygen into the bottle inthe presence of heat can lead to oxidation of ethanol into aldehydes.These processes lead to changes in odor and flavor, and thereforespoilage, of fine wines.

Current methods for identifying acetic acid in wine are very sensitive,detecting roughly 50 μ/L acetic acid, even though the accepted spoilagelimit of acetic acid in wine is 1.4 g/L (see, for example, Vonach et al.(1998) “High performance liquid chromatography with real-timeFourier-transform infrared detection for the determination ofcarbohydrates, alcohols and organic acids in wines” J. Chromatogr. A.824:159-167; Castinñeira et al. (2000) “Simultaneous determination oforganic acids in wine samples by capillary electrophoresis and UVdetection: optimization with five different background electrolytes” J.High Resol. Chromatogr. 23:647-652; Schindler et al. (1998) “A rapidautomated method for wine analysis based upon sequential injection(SI)-FTIR spectroscopy” Fresenius J. Anal. Chem. 362:130-136; andMargalith (1981) in Flavour Microbiology, pp. 167-168, Charles ThomasPublishers, Springfield, Ill.). In addition, nuclear magnetic resonance(NMR) spectroscopy has been employed for wine fingerprinting studies andtrace amino acid and organic molecule detection in wine (Guillou andReniero (1998) “Magnetic resonance sniffs out bad wine” Physics World11:22-23; and Ko

sir et al. (1998) “Wine analysis by 1D and 2D NMR spectroscopy” Analysis26:97-101). However, all published NMR studies of wine involve removaland analysis of small volume samples of wine (e.g., less then 1 mL) toaccomplish these measurements. As such, all of the current strategiesfor contaminant (e.g., acetic acid) detection require the bottle to beviolated, a process that destroys the cork, seal, and label, severelydevaluing both the wine and bottle. The present invention overcomesthese and other problems by providing methods and devices for thedetection of contaminants in wine bottles by NMR spectroscopy. Thesemethods are equally applicable to other sealed consumables containersfor which contamination, degradation, or other changes in product flavoror quality is a concern.

SUMMARY OF THE INVENTION

The present invention provides methods and devices for the analysis ofsealed consumable containers by NMR spectroscopy. The high static andradiofrequency (rf) magnetic fields used in the NMR experiment in no wayaffect the quality of the food or beverage examined via the methodsprovided herein.

In some embodiments, the present invention provides non-invasive,non-destructive analytical methods for determining the level of wineacetification. As such, the methods and devices of the present inventioncan be routinely used in the evaluation of the quality of fine wines andin the study of wine cork aging. Furthermore, these methods of intactbottle analysis are not limited to the determination of acetic acidspoilage and content in wines, but can be extended to the study of otherwine molecular components (e.g., aldehydes and flavenoids), as well asto components and/or contaminants in other types of sealed consumables.

Accordingly, the present invention provides methods for analyzing one ormore contents of a sealed consumables container. The methods include,but are not limited to, the steps of providing an NMR spectrometer andan NMR probe configured to accept a portion of the sealed consumablescontainer; positioning the portion of the sealed consumables containerwithin a data collection region of the NMR probe; establishing ahomogeneous static magnetic field across the data collection region;collecting an NMR spectrum; and analyzing one or more peaks in the NMRspectrum, thereby analyzing one or more contents of the sealedconsumables container.

Any food or beverage having components that generate one or more NMRpeaks can be assessed using the methods and devices of the presentinvention. Thus, a variety of food or beverage containers having, forexample, nonalcoholic beverages, alcoholic beverages, beer, vinegar orolive oil stored therein, can be analyzed using the methods of thepresent invention. In a preferred embodiment, the sealed consumablescontainer is a bottle of wine.

The methods of the present invention can be used in a qualitative orquantitative manner, e.g., either the presence of a selected componentor the concentration of the selected component is determined. Forexample, in the analysis of wine, exemplary selected components include,but are not limited to, acetic acid, aldehydes, flavenoids, and aminoacids.

The methods of the present invention include the step of positioning theportion of the consumables container within a data collection region ofthe NMR probe. For example, either the neck of the container or aportion of the body of the container can be placed within the datacollection region of the NMR probe. The homogeneous static magneticfield is then established across the data collection region by, forexample, adjusting the one or more shim coils in the probe. Preferably,establishing the homogeneous field allows for resolution of chemicalshift difference between selected NMR spectra peaks a minimum distanceapart. In certain embodiments of the present invention involving ¹H NMRspectroscopy, the resolution will preferably allow for distinguishingpeaks that are about 1 ppm apart. Optionally, the NMR peaks generated bythe selected components are integrated, thereby analyzing a quantity ofthe selected component.

The present invention also provides NMR probes configured to position aportion of a sealed consumables container within an NMR spectrometer.The NMR probes used in the present invention can be any of a number ofdetection probes, including, but not limited to, a ¹H probe, a ²H probe,a ¹³C probe, a ¹⁷O probe, or a combination thereof. The NMR probecomponents include a body structure having a cavity adapted forreceiving a portion of the sealed consumables container (e.g., a neck ofa bottle, or a body of the container). The cavity is typically disposedin the body structure (either at a first end, or in a middle portion),such that a first rf coil attached to the body structure is positionedproximal to the cavity and the portion of the sealed container. In oneembodiment of the probes of the present invention, the first rf coilcomprises a split solenoid coil, in which the coil portions arepositioned to either side of the data collection region of the probe. Inan alternate embodiment, the first rf coil is a birdcage-style coilsurrounding the data collection region of the probe.

In some embodiments of the present invention, the first rf coil is usedfor both transmitting and receiving rf pulses. Optionally, the probeincludes a second rf coil positioned distal to the first rf coil. Thesecond rf coils can be, for example, configured for measurement of oneor more signals from a calibration sample. Alternatively, the second rfcoil is configured for selective excitation of a heteronucleus, such as¹³C, ¹⁷O, ²H, ²³Na, ²⁷Al, ¹⁹⁹Hg, or ²⁰⁷Pb.

The probes of the present invention further include a tuning capacitorcoupled at a first position to the rf coil, and coupled at a secondposition to a length of coaxial cable configured for connection to theNMR spectrometer. The tuning capacitor can include, but is not limitedto, one or more non-magnetic zero-to-ten (0-10) picofarad high power rfcapacitors.

Optionally, the probe also includes additional components useful for NMRanalyses, such as electronic components for generating magnetic fieldgradients, a calibration fluid sample tube; and a fluid jacket formodulating the probe temperature, to name a few.

Systems for analyzing contents of a sealed consumables container arealso provided by the present invention. The system components include,but are not limited to, the NMR probe configured to position a portionof a sealed consumables container within an NMR spectrometer; an NMRspectrometer having a bore proximal to a magnet and configured toreceive the NMR probe, an amplifier coupled to the NMR probe viaco-axial cable; and a receiver system having a preamplifier and adetector. Optionally, the system further includes a pulse programmer.

Optionally, the NMR probe of the system is a single resonance probeselected from the group consisting of a ¹H probe, a ²H probe, a ¹³Cprobe, an ¹⁷O probe, a ²³Na probe, an ²⁷Al probe, a ¹⁹⁹Hg probe, and a²⁰⁷Pb probe. In one embodiment, the NMR probe employs a first rf coilused for both transmitting and receiving rf pulses. In another aspect,the NMR probe further comprises a second rf coil configured, forexample, for measurement of one or more signals from a calibrationsample.

The NMR probe is configured to accept the sealed consumables containerand position a portion of the container (e.g., the neck of a bottle, orthe body of the container) within the magnetic field of thespectrometer. Typically, the spectrometer comprises a wide bore magnet;preferably, the magnetic field is generated by a room temperaturesuperconducting magnet. While any field strength can be used in thesystem of the present invention, higher field strengths are preferableto lower field strengths. In one embodiment, magnetic field comprises a2.01 T magnetic field. The receiver component of the analytical systemincludes, but is not limited to, preamplifier and a detector incommunication with the NMR probe. In one embodiment, the receiverincludes a passive rf duplexer and signal mixing and digitizationelectronics. These and other aspects of the present invention areprovided herein.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic drawing of an exemplary probe of the presentinvention.

FIG. 2 depicts an expanded view of an exemplary probe, showing theplacement of a sealed container within the data collection region.

FIG. 3 panels A and B represent one embodiment of the systems of thepresent invention, depicting an experimental setup used to obtain an NMRspectrum of a full, intact wine bottle. FIG. 3A provides a schematicdepicting the placement of the sealed consumables container (a winebottle) and NMR probe within the body structure of an NMR spectrometer.FIG. 3B shows an expanded view of the probe, depicting the positioningof the selected portion of the container with the rf coils of the probe,and indicating that the NMR probe head is capable of housing an entirebottle of wine.

FIG. 4 panels A and B depict an alternative embodiment of the systems ofthe present invention, showing the placement of the body of the sealedconsumables container within the NMR probe. FIG. 4B shows an expandedview of the probe, depicting the positioning of the body of thecontainer within the sample measurement region of the probe.

FIG. 5 depicts NMR spectral data obtained at 9.1 T for a 500 μL sampleof wine (panel A) and red wine vinegar (panel B).

FIG. 6, panels A and B, provides spectral data generated for a sample ofwine (panel A) and a sample of red wine vinegar (panel B) using themethods and probes of the present invention.

FIG. 7 provides a plot comparing the experimental versus calculatedvalues of acetic acid provided in a set of acetic acid/ethyl alcoholfull bottle standard samples.

FIG. 8 panels A, B and C provide exemplary rf pulse sequences for use inthe methods of the present invention.

FIG. 9 panels A and B depict ¹³C NMR spectra on full bottles of wine.

FIG. 10 panels A and B are tables depicting NMR-derived percentages ofethanol (FIG. 10A) and acetic acid concentrations (FIG. 10B) in avertical series of sealed full bottles of the UC Davis CabernetSauvignon bottled between 1950 and 1977.

DETAILED DESCRIPTION

Definitions

Before describing the present invention in detail, it is to beunderstood that this invention is not limited to particular devices orcontainer systems, which can, of course, vary. It is also to beunderstood that the terminology used herein is for the purpose ofdescribing particular embodiments only, and is not intended to belimiting. As used in this specification and the appended claims, thesingular forms “a”, “an” and “the” include plural referents unless thecontent clearly dictates otherwise. Thus, for example, reference to “acapacitor” includes a combination of two or more capacitors; referenceto a “coil” includes mixtures or series of coils, and the like.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which the invention pertains. Although any methods andmaterials similar or equivalent to those described herein can be used inthe practice for testing of the present invention, the preferredmaterials and methods are described herein. In describing and claimingthe present invention, the following terminology will be used inaccordance with the definitions set out below.

The term “consumables” as used herein refers to a food, beverage, oralternate energy source (e.g., bacterial media) intended for consumptionby an organism (e.g., a human, an animal, a cell culture, and the like).Thus, the term “sealed consumables container” refers to a packaged orunopened vessel or receptacle containing the food or beverage. SealedNMR tubes prepared with samples of food or beverage products are notconsidered sealed consumables containers with respect to the presentinvention.

The terms “NMR probe” and “probe head” are used interchangeably hereinto refer to the component of an NMR spectrometer system which transmitspulses to the sample and receives the NMR signals generated.

The term “data collection region” refers to the portion of the NMR probein which the NMR signal is generated; typically, the homogeneity of themagnetic field of the spectrometer is optimized in this region.

The term “rf coil” refers to a set of filamentary wire sections arrangedin a helical geometry and designed for transmitting and/or receivingradiofrequency signals.

The term “tuning capacitor” as used herein refers to one or morecapacitor components of the NMR probe which are typically used to matchand tune the probe to the correct Larmor frequency and impedance matchthe rf circuit to, for example, 50 Ω.

The term “split solenoid coil” (or “split pair solenoid” ) refers to asolenoid having multiple coils of wire (usually in cylindrical form)that generates a magnetic field when carrying a current.

Methods

The present invention provides methods of analyzing one or more contentsof a sealed consumables container. The sealed consumables container canbe any of a number of food or beverage containers having contents ofinterest, including, but not limited to, alcoholic or nonalcoholicbeverages.

In a preferred embodiment, the container is a corked (e.g., unopened)bottle of wine. Any number of wine bottle “styles” can be accommodatedin the methods (as well as devices and systems) of the presentinvention. For example, the methods of the present invention can be usedto analyzed the contents of the high shouldered “Bordeaux” bottle(typically used for Sauvignon Blanc, Cabernet Sauvignon, Merlot, andBordeaux blends), the slope shouldered “Burgundy” bottle (Chardonnay andPinot Noir), or the taller “Hoch” bottle of Germanic origin (Rieslingsand Gewuürztraminers). The contents of champagne/sparkling wine bottles,Chianti bottles, and the shaped-neck bottles typically used forfortified wines (port, sherry, etc.) can also be analyzed by the methodsof the present invention. Furthermore, a range of bottle sizes can beused in the methods of the present invention; in addition to the 750 mLbottle found in typical wine cellars, the smaller half bottles, “splits”(187 mL) and “tenths” (375 mL) as well as the larger “magnum” bottles(e.g., 1 L, 1.5 L and 3 L bottles) can be examined.

The bottle can be made of either clear or colored (e.g., amber, green orbrown) glass. In addition, the beeswax seal and/or lead cap often usedin the corking process need not be removed for the analysis.

In addition to wine, other consumables can be analyzed by the methods ofthe present invention, including, but not limited to, beer, vinegar andolive oil. In addition, sealed receptacles containing solutions orsuspensions not typically considered as “food” (e.g., microbial culturemedia, herbal tinctures, and the like) can also be examined using themethods of the present invention. Preferably, the component of interestin the sealed container generates an NMR spectrum having at one or moresharply defined peaks.

The methods of analyzing one or more contents of the sealed consumablescontainer employ an NMR spectrometer and an NMR probe configured toaccept a portion of the sealed consumables container. In one embodimentof the present invention, the NMR probe is configured to receive thenarrowed upper portion, or “neck,” of the sealed container. In analternate embodiment, the body of the container is the portion placed inthe NMR probe.

In the methods of the present invention, the selected portion of thesealed consumables container is positioned within the data collectionregion of the NMR probe. This can be achieve by placing the containerwithin the probe, and then inserting the probe into the spectrometer,such that the selected portion of the container (neck, body, etc.) isoptimally positioned within the magnetic field of the spectrometer.Alternatively, the probe can be installed into the spectrometer prior toinsertion of the container. In either case, the container is positionedsuch that a portion of the consumables is positioned within the magneticfield of the spectrometer, and proximal to the rf coil of the NMR probe.The examined portion of the sealed container will be determined in partby the shape and/or configuration of the sealed container, as well asvarious requirements with respect to the type of NMR spectroscopyperformed. For example, either the neck of the container or a portion ofthe body of the container can be placed within the data collectionregion of the NMR probe.

In one preferred embodiment, the rf coil “examines” the neck of a winebottle between the base of the cork and the flared body of the winebottle. Although there is less sample in this region (and therefore lesssignal) as compared to the larger body of the bottle, it is easier toestablish a homogeneous static magnetic field over this smaller sampleregion, thus enhancing the probability of obtaining narrow (resolved)NMR spectral peaks.

A homogeneous static magnetic field is established across the datacollection region of the NMR probe by standard mechanisms, e.g., byadjustment of cryogenic and/or room temperature (RT) magnetic fieldshims. The NMR spectrum is then collected by monitoring the response ofthe sample to an rf electromagnetic field pulse generated by the rfcoil. Preferably, the magnetic field established is homogeneous enoughto allow for resolution of chemical shift differences between selectedNMR spectra peaks set a minimum distance apart. The degree ofhomogeneity necessary for performing the methods of the presentinvention will depend on a number of factors, including nucleiselection, magnetic field strength, and chemical structure. In themethods of the present invention, the homogeneous static magnetic fieldis established such that one or more peaks of interest from thecontaminant are resolved from additional NMR spectral peaks. Forexample, for ¹H NMR spectra collected on the contents of sealed winebottles, the minimum desired resolution is approximately 1 ppm, thedistance between the methyl resonance and the methylene resonance of theacetic acid contaminant. Exemplary NMR spectra of a number of compoundscan be found, for example, in the Aldrich Library of ¹ H and ¹³ C FT NMRSpectra, Edition 1 (1993, volumes 1-3, eds. Pouchert and Behnke, AldrichChemical Company), from which a desired minimum resolution can bereadily determined by one of skill in the art.

Since the magnetic field is not stabilized with a flux-locked loop, anda ²H lock (as typically employed with small volume NMR samples “spiked”with a deuterated standard such as TMS) is not possible for sealed winebottles, data collection is typically performed via block averaging(e.g., n data sets of free induction decay each derived from m scans).In a preferred embodiment, the data are collected as block averages ofn=10 groups of 100 scans. The n=10 free induction signals are Fouriertransformed, overlapped by shifting the frequency, and added offlineusing Matlab (Mathworks Inc, Natick Mass.). This procedure eliminatesthe effect of the long time drift in the static magnetic field on thecollected data, thereby producing highly resolved ¹H NMR spectra for themethyl group region in wine.

After an NMR spectrum is collected, the peaks of the spectrum areexamined. Typically, the analysis involves examination ofpreviously-identified peaks in a select region of the spectrum. Thepeaks can represent any of a number of components found in the sealedcontainer. For example, the peaks of interest are optionally generatedby contaminating molecular species (contaminants) indicating spoilage orexposure to oxygen. For embodiments involving the analysis of wine, oneparticular contaminant of interest is acetic acid, which is generated bythe bacterial metabolism of ethyl alcohol. For analysis of acetic acid,the regions of interest are around 1 ppm (the region in which the methylpeak for acetic acid can be found) as well as around 3.6 ppm (the regionin which the methylene peak from acetic acid is located). Alternatively,wine components such as aldehydes or flavenoids can be examined.

In some embodiments of the method, the analysis is on a qualitativelevel, e.g., are the NMR peaks of interest present or absent. In otherembodiments, the analysis is quantitative; the selected peaks areintegrated and compared to a standard peak intensity, thereby providinga quantitative analysis of the selected components of the sealedconsumables container. Preferably, the NMR resonances generated by thecomponent of interest are sharp, facilitating the optional integrationprocess. The integration can be performed using a software programprovided with the spectrometer operational software, or it can beperformed the old-fashioned way, by printing the spectra, cutting outthe peaks of interest, and weighing the paper scraps.

NMR Probes

The present invention also provides NMR probes for use in the methodsdescribed herein. The NMR probes of the present invention are configuredto position a portion of a sealed consumables container within an NMRspectrometer, thus avoiding the need to violate the seal on thecontainer in order to analyze the contents. The probes typicallycomprise a body structure having a cavity disposed at a first end of thebody structure, a first rf coil positioned proximal to the cavity andthe portion of the sealed container; and a tuning capacitor coupled tothe rf coil and to a length of coaxial cable configured for connectionto the NMR spectrometer. In an alternate embodiment, the cavity isdisposed in a middle region of the body structure, rather than proximalto the end of the probe.

The probes of the present invention can be used to detect any desirednuclei capable of generating a nuclear magnetic resonance and havingadequate chemical shift dispersion between selected contaminant and/orsample signals. Thus, the probes of the present invention include, butare not limited to, ¹H probes, ²H probes, ¹³C probes, ¹⁷O probes, andthe like. Furthermore, the probes of the present invention can be singlefrequency or dual frequency probes (e.g., a ¹H/¹³C probe).

The body of the probe is typically composed of material having a lowmagnetic susceptibility to reduce and/or prevent distortion of thestatic magnetic field when the probe is positioned in the spectrometer.Exemplary materials used in the manufacture of the body structure (orportions thereof) include, but are not limited to stainless steel,aluminum, glass, ceramic, and plastics such as Teflon(polytetrafluoroethene), Kel-F (polychlorotrifluoroetene), and PVC(polyvinylchloride).

The body structure has a cavity that is configured to accept a portionof the sealed consumables container, such that a portion of thecontainer is positioned within the data collection region of the probe.Thus, the sample cavity is greater than that typically employed in anprobe configured for NMR tubes. The overall dimensions of the probeoptionally range from about 600 mm to 800 mm in length, preferably about700 mm. The outer diameter of the probe ranges in size from 100 mm to150 mm in diameter, although an outer diameter of up to 310 mm ispossible with the current magnet embodiment. The size of the cavityportion of the probe will depend upon the sealed container to beanalyzed; for a probe configured to accept a neck portion of a winebottle (FIG. 3A and 3B), the cavity portion of the probe will typicallyrange from 34 mm to 85 mm in diameter. Larger cavities able to encompassa wider portion of a consumables container, such as the base and body ofa wine bottle (e.g., about 100-150 mm in diameter), are alsocontemplated (see FIGS. 4A and 4B).

The cavity is configured to hold the sealed container in positionthrough the use of, for example, one or more PVC positioning rings. Inone embodiment, the cavity extends from one end of the probe to the datacollection region, for insertion of the sealed container from the openend. In an alternate embodiment, the cavity is enclosed within the bodystructure, and accessed by an opening in the side of the body structure.

The first rf coil is positioned in the body structure of the probe,proximal to the cavity (and the selected region of the sealed containerinserted therein). Optionally, the first rf coil functions as both thetransmitting coil and the receiving coil. In one embodiment, the firstrf coil is a split solenoid coil. An exemplary split solenoid coil is 12gauge copper wire wound in a 1 cm diameter spiral, the first coilportion having 4 turns of the copper wire, and coupled (via a connectingportion of the wire) to a second coil portion having another 4 turns ofcopper wire. The first coil portion is positioned on one side of thecavity, while the second coil portion is positioned on an opposite sideof the cavity; the connecting wire runs between the two portions withoutcrossing the cavity itself (e.g., along the circumference of thecavity). Preferably, the second coil portioned is aligned along a sameaxis as the first coil portion.

In another embodiment, the rf coil circumscribes the cavity (e.g., thewalls of the body structure defining the cavity act as a former aroundwhich the rf coil is wound.) In a further embodiment of the probe, thefirst rf coil comprises a birdcage-style coil. Such a configuration ofcoil portions is described in, for example, Hayes et al. (1985) “Anefficient, highly homogeneous radiofrequency coil for whole-body NMRimaging at 1.5 T” J. Magn. Reson. 63:622-628.

The probes of the present invention also include one or more tuningcapacitors. The tuning capacitor is coupled at a first position to thefirst rf coil, and coupled at a second position to a length of coaxialcable configured for connection to the NMR spectrometer. In oneembodiment, the tuning capacitor is a non-magnetic 0-10 picofarad highpower rf capacitor.

A schematic representation of the probes of the present invention isshown in FIG. 1. Probe 10 comprises body structure 20, first rf(radiofrequency) coil 30; and tuning capacitors 40 and 42. Bodystructure 20 has opening or cavity 50 disposed at one end for receivingthe sealed consumables container (not shown).

A portion of cavity 50 extends into data collection region 60 of probe10. First rf coil 30 is attached to capacitor 40 at a first end 32 andattached to capacitor 42 at a second end 34, and is positioned proximalto cavity 50 such that coil portions 36 and 38 are situated to eitherside of data collection region 60.

Tuning capacitors 40 and 42 are also coupled at second positions 44 and46 to coaxial cables 70 and 72, which are configured for connection tothe NMR spectrometer (not shown). In addition, tuning capacitor 42 iscoupled at a third position to rf in/out coaxial cable 74, whichprovides the radiofrequency signal for NMR spectrum generation.

FIG. 2 depicts an expanded view of exemplary probe 110, showing theplacement of sealed container 100 within the data collection region 160.Coil portions 136 and 138 of rf coil 130 are approximately 2.0 cm indiameter (measurement A) and extend approximately 2.5 cm from the uppersurface of tuning capacitors 140 and 142, respectively (measurement B),such that the total height of rf coil 130 is approximately 4.5 cm. Coilportions 136 and 138 are positioned approximately 3.4 cm apart(measurement C) with the intermediate coil portion (represented bydotted line) arcing between the two portions, such that neck portion 102of sealed container 100 can be positioned between coil portions 136 and138 for optimal data collection. Optionally, container 100 will havestopper 104 positioned at the distal end of neck portion 102. Stopper104 is optionally a cork, a screw-top cap, or a plug. Preferably, bottle100 is positioned within data collection region 160 such that stopper104 does not interfere with the data collection procedure.

Probe 110 optionally includes positioning ring 112 separating rf coil130 from the main portion of cavity 150; the aperture in positioningring 112 allows the selected portion of bottle 100 to be positionedwithin data collection region 160 while protecting this region fromdust, etc. Optional capacitor stand 114 is positioned on the distal sideof tuning capacitors 140 and 142. Capacitors 140 and 142 areapproximately 4.5 cm in height; therefore, the distance between a faredge of coil portion 136 and the distal side of capacitor 140 isapproximately 9 cm, and the distance between outer edges of positioningring 112 and capacitor stand 114 is approximately 11 cm.

The probes of the present invention optionally incorporate a second rfcoil, preferably positioned distal to the first rf coil. The second rfcoil can be employed for a number of purposes. For example, the secondrf coil can be used for either transmitting or receiving the rf signal(in embodiments in which the first rf coil does not function as bothtransmitter and receiver). Alternatively, the second rf coil can beconfigured for measurement of one or more signals from a calibrationsample. In yet another embodiment, the second rf coil provides forselective excitation of a heteronucleus (including, but not limited to,¹³C, ¹⁷O, ²H, ²³Na, ²⁷Al, ¹⁹⁹Hg, ²⁰⁷Pb, and the like).

Optionally, the probe further includes one or more components for tuningand/or impedance matching the rf coil(s) to at least one rf power sourceat a selected frequency.

The probes of the present invention optionally include one or moreadditional components which enhance the functioning of the probe. Forexample, the probe can include components for generating magnetic fieldgradients, which can be used, for example, for imaging purposes. In someembodiments, the probe includes a calibration fluid sample tube. Theoptional calibration sample tube is typically positioned within thecavity of the body structure such that the calibration sample ispositioned proximal to the selected portion of the sealed consumablescontainer when the container is inserted in the cavity.

In a further embodiment, the NMR probes of the present inventionoptionally further include a fluid jacket, reservoir or other mechanismfor modulating the temperature of the probe. Exemplary fluid jacketdesigns for use with the present invention are described in, forexample, U.S. Pat. No. 5,530,353 titled “Variable Temperature NMR Probe”(Blanz).

System Components

The present invention also provides systems for analyzing contents of asealed consumables container. The systems include one or more NMR probesof the present invention, an NMR spectrometer, and a receiver systemconfigured for electronic communication with the NMR probe. The probesand systems of the present invention can be used to perform pulsed,continuous wave, or gradient NMR experiments.

The NMR spectrometer typically comprises a body structure, a magnethoused within the body structure, a bore proximal to the magnet andconfigured to receive the NMR probe, and an amplifier configured forcoupling to a first position on the NMR probe. Optionally, the magnet isa constant external magnet, a room temperature (RT) magnet, and/or asuperconducting magnet. Any NMR spectrometer having a bore capable ofreceiving the NMR probes can be used in the systems of the presentinvention. Preferably, the NMR spectrometer is a super wide borespectrometer. Exemplary spectrometers are available commercially from,for example, Varian (Palo Alto, Calif.; www.varianinc.com) and Bruker(Germany, www.bruker.com). The field strength of the magnet componentused in the systems can also vary, ranging from 2.01 T to 9.4 T andhigher.

The systems of the present invention include a receiver systemconfigured for electronic communication with the NMR probe. Optionally,the receiver system is incorporated into the body structure of the NMRspectrometer. The receiver system typically comprises a preamplifierconfigured for coupling to the NMR probe and a detector in communicationwith the preamplifier. In one embodiment of the systems of the presentinvention, the receiver includes a passive rf duplexer as well aselectronics for signal mixing and digitization (see, for example,Fukushima and Roeder, Experimental Pulse NMR a Nuts and Bolts Approach,Addison-Wesley, New York, 1981).

Optionally, the system further includes an NMR pulse programmer.Exemplary pulse programmers are available from Tecmag, Inc. (Houston,Tex.; www.tecmag.com).

In some embodiments of the present invention, the system includes amechanism for spinning the sealed container within the NMR probe.Exemplary spinning mechanisms include, but are not limited toair-propelled mechanisms (e.g., air turbines), rotor mechanisms,strap-based mechanisms and the like.

In a preferred embodiment of the present invention, the system alsoincludes a rf power source, for exciting the nuclei within the sealedcontainer.

FIG. 3A provides an exemplary system of the present invention depictingthe positioning of bottle 200 within the data collection region 260 ofprobe 210, which is inserted into magnet 280 of the NMR spectrometer.FIG. 3B shows the alignment of bottle 200 within probe 210 with respectto rf coil 230 and tuning capacitors 240 and 242. Also depicted areoptional components positioning ring 212 and capacitor stand 214.

FIGS. 4A and 4B depict an alternate positioning of bottle 300 within thedata collection region of probe 310, in which the body of bottle 300 isinserted into data collection region 360. In FIG. 4A, probe 310 isinserted into magnet 380 of the NMR spectrometer. FIG. 4B shows rf coil330, tuning capacitors 340 and 342, coaxial cables 370 and 372, and rfin/out cable 374, with respect to the alignment of bottle 300 withinprobe 310. Positioning ring 376 centers the sample inside of rf coil330, which is mounted on PVC positioning rings 378 and 379.

EXAMPLES

It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be included within the spirit and purview of this application andscope of the appended claims. Thus, the following examples are offeredto illustrate, but not to limit the claimed invention.

The methods, NMR probes and spectrometer systems of the presentinvention are capable of detecting less then 0.5 g/L amounts of aceticacid in wine. For the analysis of acetic acid content, the acetic acidmethyl group hydrogen nuclei and the ethyl alcohol methyl group hydrogennuclei are examined, which differ in chemical shift by approximately 1ppm. This method for acetic acid quantitation does not violate the winebottle, is harmless to the bottle contents, and can be easily extendedto the exploration of other vital ingredients and or contaminants inintact wine bottles and other sealed consumable containers.

Example 1

Determination of Acetic Acid Levels in Wine Samples

Standards Preparation

The titration experiments were performed on full bottle acetic acidstandards prepared from mixtures of de-ionized water, 200 proof ethylalcohol obtained from Gold Shield Chemical Co. (Hayward, Calif.), and99.7% glacial acetic acid purchased from EM Science (Gibbstown, N.J.).The control samples were generated by filling, or “charging” empty winebottles with 750 mL of 12.5% (v/v) ethyl alcohol in water having aselected concentration of acetic acid (ranging between about 0.5 g/L andabout 3.2 g/L). Sodium chloride (Fisher Scientific, Hampton, N.H.) wasdissolved in 750 mL water and used as a calibration standard for bothshimming the magnetic field for nuclei with low gyromagentic ratio γ andfor determining the Larmor frequency of the comparatively less sensitive¹³C nucleus in full bottle wine samples. The tested wines were eitherpurchased from local markets or obtained as gifts from the UC DavisDepartment of Viticulture and Enology.

Experimental Set-up

The NMR experiments on sealed wine bottles were performed at 2.01 Tmagnetic fields corresponding to a ¹H Larmor frequency of 85.78 MHzrespectively. A high field NMR spectrometer (Varian Inc. Inova 400, PaloAlto, Calif.) employing a 9.1 T magnetic field (corresponding to a HLarmor frequency of 399.76 MHz. The ) was used to confirm the aceticacid concentrations measured on the low field instrument, using 500 μLaliquots of the samples.

The single resonance NMR spectrometer delivers rf pulses to a high poweramplifier connected to the NMR probe head mounted inside of an OxfordInstruments (Palo Alto, Calif.) 310 mm room temperature boresuperconducting solenoid imaging magnet. The full intact wine bottle ishoused inside of the NMR probe head as shown in FIG. 3A. The rf coil isproximal to the neck of the wine bottle between the base of the cork andthe main body of the wine bottle.

Careful adjustment of the cryogenic and room temperature magnetic fieldshims was performed to establish a homogeneous field over the winebottle (as indicated by a ¹H line width of ≦4 Hz). Although there isless sample in this region of the bottle in comparison to the bottlebody and base, it is much easier to establish a homogeneous staticmagnetic field over the small sample region and ultimately producenarrow, highly resolved NMR lines. Current for the room temperatureshims was provided by a General Electric Omega series NMR spectrometermagnetic field shim power supply, modified to output −5 V to +5 V DC oneach channel. The supply was controlled by a potentiometer bank obtainedfrom a Varian EM 390 (90 MHz) continuous wave NMR spectrometer.

After termination of the rf pulse, the sample emits a low μV-mV rfsignal that is mixed to audio frequencies and digitized by the NMRspectrometer. Fourier transformation of this signal yields the standardNMR spectrum. Substantial improvements in dynamic range can be made byselectively exciting and measuring just the methyl group region of the¹H NMR spectrum between 1 and 2.5 ppm. Operation in this way removes themassive background signal from water at 4.8 ppm.

Several different nuclei including deuterium (²H), oxygen (¹⁷O), carbon(¹³C), and hydrogen (¹H) were considered as possible candidates fordetermining acetic acid levels in wine. Of these possibilities, the ¹Hnucleus was chosen due to its superior sensitivity and the 1 ppmchemical shift difference between the spectrum of acetic acid and thespectra of water and ethyl alcohol, the two major constituents of wine.

NMR Data Collection: Control Data

The presence and/or quantity of acetic acid in various wine-basedsamples was determined by ¹H NMR spectroscopy at 9.1 T as a control.FIG. 5A provides a portion of an NMR spectrum generated for a 500 μLsample of the 1997 vintage UC Davis Experimental Vineyard CabernetSauvignon. The intense peak at 4.8 ppm is due to water, while thequartet and triplet centered at 3.6 ppm and 1.1 ppm represent themethylene and methyl groups in ethyl alcohol, respectively. The ¹H NMRspectrum in FIG. 5B (also obtained at 9.1 T) corresponds to a homemadesample of red wine vinegar. The new peak at approximately 2 ppm clearlyindicates the methyl group in acetic acid, and the lack of splittings ofthis single line is consistent with the chemical structure. The amountof acetic acid in the red wine vinegar was determined to be 2.6% (or27.6 g/L), based upon the ratio of the methyl group peak heights in FIG.5B, assuming that the ethyl alcohol was 12.5% of the full bottle volumeprior to acetification.

NMR Data Collection: Experimental Data

Exemplary data collected by the methods and probes of the presentinvention is shown in FIGS. 6A and 6B. The ¹H NMR spectrum in FIG. 6Awas obtained for a full bottle of the UC Davis Cabernet Sauvignon, withselective excitation of the methyl group frequencies between ±3 ppm. Thetriplet-splitting of the methyl resonance depicted in FIG. 6A(corresponding to the triplet shown at 1.1 ppm in the 500 μL sample ofFIG. 5A) is due to scalar coupling with the protons in the methylenegroup in the ethyl alcohol molecule. The full bottle ¹H NMR spectrumshown in FIG. 6B, corresponds to a 750 mL mixture of water, 12.5% ethylalcohol, and 0.5% acetic acid. The singlet peak centered at 2.1 ppm(present in the FIG. 6B vinegar sample but not the FIG. 6A wine sample)clearly indicates the presence of acetic acid (as expected) fromcomparison to the spectrum obtained for the small volume shown in FIG.5B. The NMR spectrum in FIG. 6B corresponds to an acetic acidconcentration of 5.3 g/L, nearly 3.8 times the accepted 1.4 g/L aceticacid spoilage limit for wine.

Titration Data

The titration data shown in FIG. 7 provides a comparison of the preparedacetic acid concentrations versus NMR measurements of acetic acidconcentration in the prepared samples, as determined from the ratio ofthe integrated area of the acetic acid peak at 2.1 ppm to the integratedarea of the ethyl alcohol triplet at 1.1 ppm given the 12.5% (v/v) ethylalcohol concentration. The open circles correspond to the average ofnine measurements of the acetic acid concentration from full bottle NMRspectra at 2.01 T, while the open triangles represent one measurement ofthe acetic acid concentration in a 500 μL sample at 9.1 T. The dashedline of unit slope is included in FIG. 7 indicate the correlationbetween prepared and experimentally-determined concentrations of aceticacid. Both the low field “full bottle” measurements and the high field“small sample” measurements of acetic acid agree with preparedconcentrations, although there is some spread in the data. In the caseof the high field small sample results, the uncertainty between theprepared and measured concentrations is most likely due to a liquidvolume measurement error in the sample preparation, as the extremelynarrow ¹H NMR line widths as shown in FIG. 5 permit reasonably accuratepeak intensity calculation by integration. The increased line widths inthe full bottle experiment shown in FIG. 6 introduce more error into themeasurement of acetic acid concentration as shown by the error bars inFIG. 7, due to the increased difficulty in assigning starting and endingpoints for peak integration. Consequently errors in both liquid volumemeasurements during sample preparation and peak intensity determinationintroduce slightly deviations from exact agreement with the dashed linein FIG. 7. Improved magnetic field shims yielding narrower lines willsubstantially increase the accuracy of the acetic acid concentration asmeasured. However, despite this small disparity, the full bottle methodis capable of evaluating the amount of wine acetification down to atleast 0.5 g/L, more than half the accepted spoilage limit of 1.4 g/L.

Further Calculations

Even though wine is an extremely complex mixture of diverse chemicalconstituents, a wine sample produces a relatively simple ¹H NMRspectrum. In the absence of spoilage, the ¹H NMR spectrum of a sample ofwine (as obtained following a single pulse excitation using the sequenceprovided in FIG. 8A) has a singlet resonance positioned at 4.8 ppm(corresponding to water), as well as an ethanol-derived quartetresonance and triplet resonance centered at 3.6 ppm and 1.1 ppm,respectively. The presence of low levels of acetic acid due to winespoilage is indicated by another singlet resonance, positioned at 2.1ppm. Taking the ratio of the integrated intensities of the ethanoltriplet to the water peak, and the acetic acid peak to the ethanoltriplet allows the percentage of ethanol by volume and the concentrationof acetic acid in wine to be quantified as: and${{EtOH}\quad\%\left( {v/v} \right)} = \frac{f_{EtOH} \times 10^{3}}{{\left( {8.5 + {8.2f_{HOAc}}} \right)f_{EtOH}} + 4.6}$${{{and}\lbrack{HOAc}\rbrack}\left( {g/L} \right)} = \frac{f_{HOAc}f_{EtOH} \times 10^{4}}{{\left( {8.3 + {8.0f_{HOAc}}} \right)f_{EtOH}} + 4.5}$where the molecular weights and densities of water, ethanol and aceticacid have been used to calculate the values in the denominator of theequations. The measurement of f_(EtOH) is derived from data collected bya one pulse experiment as depicted in FIG. 8A. However, a similarestimate of f_(HOAc) is complicated by the strong water and ethanolsignals (e.g., 99% of the spectral intensity). Since the methyl groupresonance for ethanol and acetic acid are centered at 1.1 ppm and 2.1ppm respectively, and that the water resonance is shifted 2.7 ppmdownfield from the acetic acid peak (e.g., a 232 Hz downfield shift at2.01 T), the pulse sequence provided in FIG. 8B can optionally be usedfor data generation. The combination of selective excitation, delayedacquisition and block averaging can be used reliably and reproducibly tomeasure f_(HOAc) (see Weekley et al. (March 2003) “Using NMR to studyfull intact wine bottles” J. Magn. Reson. 161:91-98). The 3 ms soft rfpulse “tips” the water magnetization by less than 5 degrees, and whencombined with a 200 Hz audio filter bandwidth, the signal intensity ofthe water peak is attenuated about an order of magnitude. The delayedacquisition combined with the long spin-spin relaxation times for themethyl protons in ethanol and acetic acid reduces the short-lived freeinduction decay (fid) components that lead to broad spectral lines, thusyielding the desired narrow resonances (e.g., line widths of approx. 4Hz).

The methyl group region of the ¹H NMR spectrum for a full bottle of 1997vintage UC Davis Cabernet Sauvignon is shown in FIG. 6A, while thecomparable data for a full bottle having 12.5% (w/v) ethanol dissolvedinto water, with 0.5% (v/v) added acetic acid is shown in FIG. 6B. Thetriplets in these spectra correspond to the ethanol methyl group, basedupon both the 1.1 ppm chemical shift and the splitting pattern (due toscalar coupling with the two equivalent methylene ¹H nuclei in theethanol structure). The single peak at 2.1 ppm in the spectrum shown inFIG. 6B corresponds to acetic acid. Using the formulas provided above,f_(EtOH) is determined to be 6.4×10⁻². The ratio of the integratedintensity of the acetic acid peak to the ethanol triplet in FIG. 6Bgives f_(HOAc) as 4.5×10⁻², which can be used to calculate that theconcentration of acetic acid [HOAc] in the sample is 5.7 g/L, ascompared to the solution as prepared (5.3 g/L of acetic acid in the 0.5%(v/v) standard solution). The 0.4 g/L difference between thesemeasurements is probably due to error in the standard preparation.

Example 2

Determining Acetic Acid Spoilage in Unopened Bottles of Wine

In most practical applications, there is no prior knowledge of theration f_(EtOH), because wines of different vintages, sources, types andquality can differ in ethanol concentration between about 7% to 24%(v/v). In these situations, the pulse sequence as provided in FIG. 8A isfirst used to measure the entire ¹H NMR spectrum, followed byapplication of the pulse sequence of FIG. 8B to selectively excite anddetect the methyl group region. In this manner, both f_(EtOH) andf_(HOAc) can be measured peak integrals and used to calculate thepercentage of ethanol and concentration of acetic acid. As noted above,data collection is typically performed via block averaging (e.g., asblock averages of n=10 groups of 100 scans. The sets of free inductionsignals are Fourier transformed, overlapped by shifting the frequency,and added offline. This procedure eliminates the effect of the long timedrift in the static magnetic field on the collected data, therebyproducing highly resolved ¹H NMR spectra for the methyl group region inwine, which can be used to accurately measure f_(HOAc).

The accuracy and sensitivity of this approach has been tested in fullbottles by comparing the NMR-derived concentrations to actual preparedconcentrations. The one-to-one agreement between the differentconcentration measurements with the less than 0.1 g/L acetic acidsensitivity of the full bottle NMR approach prompts further analysis.The NMR-derived percentages of ethanol (FIG. 10A) and acetic acidconcentrations (FIG. 10B) in a vertical series of sealed full bottles ofthe UC Davis Cabernet Sauvignon bottled between 1950 and 1977 werecompared. As expected, the amount of ethanol in this series does notcorrelate well with the year, and varies between 10-20%. Interestingly,the two most recent vintages display concentrations of ethanol veryclose to the industry standard for most wines (12.5% v/v). A similarlack of correlation is observed (FIG. 10B) for the full bottle aceticacid concentrations for these same wines. Although he oldest winedisplays the largest degree of acetic acid spoilage (6.3 g/L), and theyoungest wine has no measurable acetic acid contamination, the acidconcentration in the other vintages caries between 0.4 g/L and 2.0 g/L.It is therefore incorrect to assume that older wines will automaticallyhave a higher concentration of acetic acid as compared to younger wines.The integrity of the cork (and hence the quality of the bottle sealagainst oxygen leakage with time) is of paramount importance to aceticacid contamination.

It should be emphasized that the apparatus is capable of investigating avariety of common bottle shapes and sizes, as well as other sealedconsumables containers. All of these factors including the effects oflead or metallic seals can be compensated for by carefully adjusting thehome built room temperature magnetic field shims. Additionally, the leador metallic seals do not measurably interfere with the probe tuning orthe homogeneity and intensity of the rf field across the wine bottle.Although the titration data shown in FIG. 7 only documents results downto 0.5 g/L acetic acid, levels down to 0.1 g/L have been measured withthe probes and systems of the present invention. It is anticipated thatNMR solvent suppression techniques and/or a dual coil NMR probe headwill extend the sensitivity by one or more orders of magnitude.

Example 3

¹³C NMR Spectroscopy of Full Bottle Samples

As noted herein, the present invention for the NMR analysis of sealedconsumables containers are not limited to methods and devices involvingperforming ¹H NMR spectroscopy. In an effort to increase the sensitivityof measurements of dilute components (like flavenoids and aldehydes), aswell as to extend the full bottle technique to nuclei other than ¹H, anadditional probe embodiment was constructed (see FIGS. 4A and 4B).Instead of examining the approximately 25 cm³ sample volume in the neckof the wine bottle, the probe can be used to analyze the much larger (˜1L) volume in the body of the wine bottle. Although the magnetic fieldhomogeneity is worse across a larger sample volume, examination ofnuclei having a larger chemical shift dispersion than ¹H will be lesssensitive to the increased line width.

In one embodiment of the methods of the present invention, sealedconsumables containers are examined using ¹³C NMR spectroscopy. The muchwider chemical shift range and lower Larmor frequency of ¹³C as comparedto ¹H (21.56 MHz versus 85.78 MHz at 2.01 T, respectively) reduced thenecessity for narrow line width for analysis. As such, it becomesfeasible to center the rf detection coil on the main body of the winebottle, thereby improving sensitivity (due to greater volume of nuclei)without sacrificing the rf coil filling factor.

The formation of spin echoes for low γ nuclei is possible using theprobes of the present invention (see, for example, FIGS. 4A and 4B),despite the observation that the geometry of the four turn splitsolenoid rf coil is not optimized for homogeneity. In the special caseof ¹³C NMR spectroscopy, in which the spin-lattice and spin-spinrelaxation times tend to be long, multiple π pulse sequences (asdepicted in FIG. 8C) can be employed to refocus the magnetization andincrease the signal to noise ratio (S/N) for a fixed number of scans byadding (offline) the free induction signal following the 100 μs π/2pulse to the echo signals appearing at 102 ms intervals. In this manner,fully ¹H-coupled ¹³C NMR spectra corresponding to 100-1000 scans can beobtained for full bottle samples in a reasonable period of time

FIGS. 9A and 9B depict ¹³C spectra on full bottles of either the 1997 UCDavis Cabernet Sauvignon (9A) or red wine vinegar (9B), using the pulsesequence provided in FIG. 8C with n=7. The triplet and quartet centeredat 57 ppm and 18 ppm arise from the methylene and methyl carbons ofethanol, respectively. The line splitting of about 140 Hz in both ofthese peaks, as well as their splitting patterns, are consistent withscalar coupling to directly bonded ¹H nuclei. In the vinegar sample,additional ¹³C peaks are seen at 18 ppm and 21 ppm, due to the carbonyland methyl groups of the acetic acid. The inverted triangles in FIG. 10Blabel the acetic acid methyl group quartet. The near-equal integratedintensity of the nested quartets suggests that the amount of ethanol andacetic acid in the sample of red wine vinegar are nearly equal, a resultconsistent with the literature (Jakish (1985) Modern Winemaking CornellUniversity Press, Ithaca N.Y.).

It is clear from the spectra that the full bottle ¹³C NMR method isfeasible for the exploration of additional wine components, such astannins, flavenoids, phenols, aldehydes and amino acids. In principle,continues signal averaging will reveal these peaks in the ¹³C spectrum,although the spectra will be very complicated in the absence ofdecoupling from the ¹H nuclei. Optionally, an additional ¹H channel isincorporated into the probes of the present invention, thereby providingincreased resolution and sensitivity (and potentially, nuclearOverhauser effects) through the use of ¹H decoupling. Furthermore, probeembodiments for detection of additional isotopes, such as ²⁰⁷Pb, ¹⁹⁹Hg,⁴⁵Sc, ³⁹K, ²⁷Al, ²³Na and the like are also contemplated. Although theabundance of these isotopes is typically below the detection limit forstandard (i.e., microliter volume) NMR spectroscopy, the increasedvolumes employed in the full bottle spectroscopic methods and probesamplifies the number of spins by a factor of 10⁴, thus making the studyof trace elements in native wine samples accessible for the first time.Moreover, the methods and devices of the present invention can be usedto analyze the quality and nature of the wine bottle itself (e.g., by acombination of ²⁹Si and ²³Na NMR spectroscopy), while the cork (eithernatural or synthetic) could be studied, e.g., using ¹³C solid state NMRtechniques.

The discussion above is generally applicable to the aspects andembodiments of the present invention. Moreover, modifications can bemade to the methods, apparatus, and systems described herein withoutdeparting from the spirit and scope of the invention as claimed, and theinvention can be put to a number of different uses including thefollowing:

The use of an NMR probe configured to accept a sealed consumablescontainer or an NMR system as set for the herein, for performing any ofthe methods and assays set forth herein.

The use of an NMR probe or system as described herein for performingnoninvasive analysis of a corked wine bottle or any other sealedconsumables container, e.g., for analysis of one or more contaminants,as set forth herein.

A kit comprising one or more standard solutions of contaminant (e.g.,acetic acid titration samples) in a sealed consumables container, foruse in the methods, devices or systems of the present invention.Optionally, the kit further comprises an instruction manual forperforming the methods of the present invention.

While the foregoing invention has been described in some detail forpurposes of clarity and understanding, it will be clear to one skilledin the art from a reading of this disclosure that various changes inform and detail can be made without departing from the true scope of theinvention. For example, all the techniques and apparatus described abovecan be used in various combinations. All publications, patents, patentapplications, and/or other documents cited in this application areincorporated by reference in their entirety for all purposes to the sameextent as if each individual publication, patent, patent application,and/or other document were individually indicated to be incorporated byreference for all purposes.

1-43. (canceled)
 44. A method of assessing an anaerobic environment of asealed consumables container, the method comprising: positioning aportion of the sealed consumables container within a data collectionregion of an NMR probe configured to accept the container; establishinga homogeneous static magnetic field across the data collection region;collecting NMR data and generating a Fourier-transformed NMR spectrum;and analyzing the NMR spectrum for the presence of a contamination ordegradation component, thereby assessing the anaerobic environment ofthe sealed consumables container.
 45. The method of claim 44, whereinthe sealed consumables container is a beverage container.
 46. The methodof claim 44, wherein the sealed consumables container is a bottlecontaining olive oil.
 47. The method of claim 44, wherein positioningthe portion of the container comprises placing a neck of the containerwithin the data collection region of the NMR probe.
 48. The method ofclaim 44, wherein positioning the portion of the container comprisesplacing a body of the container within the data collection region of theNMR probe.
 49. The method of claim 44, wherein analyzing the NMRspectrum further comprises determining a concentration of thecontamination or degradation component.
 50. The method of claim 44,wherein the contamination or degradation component comprises an aceticacid.
 51. The method of claim 44, wherein the contamination ordegradation component comprises an aldehyde.
 52. The method of claim 44,wherein the contamination or degradation component comprises one or moreflavenoids.
 53. The method of claim 44, wherein collecting the NMR datacomprises attenuation of a water resonance using a radiofrequency (rf)pulse prior to data collection.
 54. The method of claim 44, whereincollecting the NMR data comprises block averaging of data sets.
 55. Themethod of claim 44, wherein establishing the homogeneous static magneticfield across the data collection region comprises adjusting one or moreshim coils.
 56. The method of claim 55, wherein adjusting the one ormore shim coils produces a ¹H line width of ≦4 Hz.